Abstract

Background

The zebrafish has been established as the main vertebrate model system for whole organism
screening applications. However, the lack of consistent positioning of zebrafish embryos
within wells of microtiter plates remains an obstacle for the comparative analysis
of images acquired in automated screening assays. While technical solutions to the
orientation problem exist, dissemination is often hindered by the lack of simple and
inexpensive ways of distributing and duplicating tools.

Results

Here, we provide a cost effective method for the production of 96-well plate compatible
zebrafish orientation tools using a desktop 3D printer. The printed tools enable the
positioning and orientation of zebrafish embryos within cavities formed in agarose.
Their applicability is demonstrated by acquiring lateral and dorsal views of zebrafish
embryos arrayed within microtiter plates using an automated screening microscope.
This enables the consistent visualization of morphological phenotypes and reporter
gene expression patterns.

Conclusions

The designs are refined versions of previously demonstrated devices with added functionality
and strongly reduced production costs. All corresponding 3D models are freely available
and digital design can be easily shared electronically. In combination with the increasingly
widespread usage of 3D printers, this provides access to the developed tools to a
wide range of zebrafish users. Finally, the design files can serve as templates for
other additive and subtractive fabrication methods.